Detection of Trichomonaisis in Vaginal and Urine Specimens from Women by Culture and PCR

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Detection of Trichomonaisis in Vaginal and Urine
Specimensfrom Women by Culture and PCR

• Mariska Petersen

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Trichomonaisis

• History
• Biological Mechanism
• Experimentation
• Results

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Trichomonaisis

• Sexually transmitted infection
• Discovered in 1836
• 5 million cases reported every year
• Resides in genitourinary tract of males and
  females
• 50 asymptomatic carriers

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Transmission

• Sexual intercourse
• Mutual masturbation
• Sharing sex toys
• Mother to child during vaginal delivery

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Signs and Symptoms

• In females there is
• inflammation of the vaginal canal
• vulvar itching leading to edema
• tenderness and chaffing
• redness
• yellow and green discharge
• painful urination
• pain during sex
• punctuate hemorrhages on the cervix known as
  colpitis macularis or strawberry cervix.

• In males there is
• whitish discharge although rare
• pain and burning during urination

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Treatment
Metronidazole (Flagyl) single two gram dose
(85-92 cure rate) OR 250 mg three time daily for
7-10 days. Tinidazole is an alternate drug
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Biological Mechanism

• T. vaginalis causitive agent
• Reproduction- binary fission
• Incubation period-4-28 days
• Facultative anerobic parasite
• Adhesion proteins on surface of flagella
• Ligand/Receptor Cytoadherence

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Experiment

• Screening for Trichomonas vaginalis by
  wet-preparation microscopy and culture.
• Presence of T. vaginalis DNA by specific PCR of
  vaginal and urine specimens.

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• Wet preparation microscopy-
• PCR-

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Materials and Methods

• Female patients from the Jefferson County
  Department of Health STD clinic were screened for
  trichomonaisis.
• Routine pelvic examinations were performed
• Swab specimens collected
• Urine samples examined

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Initial Testing

• 1st swab was used to inoculate culture medium for
  T. vaginalis
• 2nd swab placed in cryogenic airtight vial for
  PCR studies
• Wet preps were examined as a part of the routine
  exam
• All vaginal symptoms were recorded
• Urine samples were pelleted () at 1000 g for 5
  minutes, decanted and re-suspended in 250 mL of
  sterile water.
• Culture were examined daily for 5 days for the
  presence of motile trichomonads.

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PCR Testing

• Vaginal swabs agitated in 1mL of water then
  centrifuged at 2,000 3 g for 10 min.
• DNA extracted

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Results

• Samples were recorded from 190 women
• Overall presence of trichomonas was 28
• Culture of vaginal fluid most sensitive method
  for the detection of T. vaginalis- 94.3
• Direct microscopy least sensitive-58.5
• PCR-based detection of T. vaginalis from vaginal
  swabs was equivalent to culture

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Results contd

• In 74.5 of women motile trichomonads were
  detected in urine samples.
• PCR for the urine specimens detected 68.4 of
  motile trichomonads.
• 75 of women with trichomonads complained of
  symptoms including discharge, odor and itching.

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Conclusion

• T. vaginalis was detected in the urine of 75 of
  women with trichomoniasis using standard methods
• PCR of urine for T. vaginalis had a sensitivity
  less than that of microscopy and culture
• PCR for T. vaginalis using vaginal swab specimens
  was equivalent to culture

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References
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Questions???